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PHLP2 is essential and plays a role in ciliogenesis and microtubule assembly in Tetrahymena thermophila
Author(s) -
Bregier Cezary,
KrzemieŃOjak Lucja,
WŁOga Dorota,
JerkaDziadosz Maria,
Joachimiak Ewa,
Batko Katarzyna,
Filipiuk Iwona,
ŚMietanka Urszula,
Gaertig Jacek,
Fabczak StanisŁAw,
Fabczak Hanna
Publication year - 2013
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.24384
Subject(s) - tetrahymena , ciliogenesis , cilium , microbiology and biotechnology , basal body , microtubule , biology , tubulin , green fluorescent protein , cytosol , flagellum , biochemistry , gene , enzyme
Recent studies have implicated the phosducin‐like protein‐2 (PHLP2) in regulation of CCT, a chaperonin whose activity is essential for folding of tubulin and actin. However, the exact molecular function of PHLP2 is unclear. Here we investigate the significance of PHLP2 in a ciliated unicellular model, Tetrahymena thermophila , by deleting its single homolog, Phlp2p. Cells lacking Phlp2p became larger and died within 96 h. Overexpressed Phlp2p‐HA localized to cilia, basal bodies, and cytosol without an obvious change in the phenotype. Despite similar localization, overexpressed GFP‐Phlp2p caused a dominant‐negative effect. Cells overproducing GFP‐Phlp2p had decreased rates of proliferation, motility and phagocytosis, as compared to wild type cells or cells overproducing a non‐tagged Phlp2p. Growing GFP‐Phlp2p‐overexpressing cells had fewer cilia and, when deciliated, failed to regenerate cilia, indicating defects in cilia assembly. Paclitaxel‐treated GFP‐Phlp2p cells failed to elongate cilia, indicating a change in the microtubules dynamics. The pattern of ciliary and cytosolic tubulin isoforms on 2D gels differed between wild type and GFP‐Phlp2p‐overexpressing cells. Thus, in Tetrahymena , PhLP2 is essential and under specific experimental conditions its activity affects tubulin and microtubule‐dependent functions including cilia assembly. J. Cell. Physiol. 228: 2175–2189, 2013. © 2013 Wiley Periodicals, Inc.